The pregnancy-specific glycoprotein (PSG) gene family consists of 11 closely related genes that form a subgroup of the carcinoembryonic antigen (CEA) gene family on 19q13.2. Using a high-resolution restriction fragment fingerprinting technique, we have assembled 256 cosmids from the PSG region into a single 700-kb contig. Fluorescence in situ hybridization to sperm pronuclei and cosmid walking experiments indicated that this PSG contig was directly telomeric of CGM8 at the telomeric end of the CEA subgroup gene cluster. Detailed restriction mapping and hybridization with gene-specific probes indicated that the order of the 11 Previously identified PSG genes is cen-PSG3-PSG8-PSG12-PSG1-PSG6-PSG7-+ ++PSG13-PSG2-PSG5-PSG4-PSG11-tel. The CEA subgroup gene CGM11 is located at the telomeric end of the PSG gene cluster. The PSG genes are all oriented in tandem with the 5'-3' direction of transcription from telomere to centromere. The detailed map also led to the identification of seven new CEA family genes in this region. One of these (CGM12), located between CGM8 and PSG3, is a member of the CEA subgroup. The remaining six (CGM13 through CGM18) are interspersed among the PSG genes and appear to form a third distinct subgroup within the CEA gene family.